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Chemical Mechanisms of Digestion Lab

Modified from:
McGraw Hill Chemical Aspects of Digestion
Penguin Professor Digestive Enzyme Lab
Indiana State University Digestive Physiology Lab
Names of Group Members:

Safety:
DO NOT ingest any of the components.
Safety glasses should be worn at all times.
Hydrochloric acid is highly corrosive. Use extreme caution.
Materials:

16 test tubes
Plastic Cup
Transfer pipette
Hot Plate
Beaker for water bath
Water
Amylase Solution
Pepsin Solution
HCl Solution
Heavy Cream
Egg White
Starch Suspension
Lugols Reagent
pH test strips
Thermometer
Graduated cylinders

Part 1: Digestion of starch with amylase


Background: Carbohydrate digestion begins in the mouth through the break down of
starches into maltose.
In the presence of amylase
Starch + Water Maltose
Procedure:
1. Label 4 clean test tubes 1 4.
2. Obtain 10 ml of saliva (use a cup and then transfer using a disposable 1.0 ml
transfer pipette.) Think about chocolate chip cookies if necessary. If this doesnt
work, chew a rubberband. ONLY 1 PERSON PROVIDES SALIVA. NO MIXING!!!
This is your amylase solution.
3.

Obtain four test tubes and label them 1-4

4.

Add the following solutions to the four tubes (see Fig 11.7):

TUBE 1: 3.0 ml water

TUBE 2: 3.0 ml amylase solution (saliva)

TUBE 3: 3.0 ml amylase solution and 10 drops of HCl

TUBE 4: 3.0 ml amylase solution, then place tube in boiling water for 5 min

5.

Add 5.0 ml of starch solution to each tube

6.

Place all four tubes in a 37 C water bath for 1 hr to incubate

7.

After incubation, obtain another set of four test tubes labeled 1-4. Split the four
incubated solutions evenly among the two sets of tubes. One set of solutions will
be tested for the presence of undigested starch (Lugols test), whereas the other
will be tested for the presence of maltose (Benedicts test)

8.

Lugols Test (Fig 11.8) add a few drops of Lugols iodine solution to each of the
four tubes in one set of solutions.

If the solution has an amber coloration(like the Lugols reagent) then there is
no starch present (-)


9.

If the solution turns blue or black, starch is present in the solution (+)

Benedicts Test (Fig 11.9) add 5 ml of Benedicts reagent to each of the four
solutions, and place the four tubes in boiling water for 10 min.

If the solution is blue (like the Benedicts reagent), there is no maltose


present in the solution (-)

If the solution has a greenish coloration, then a small amount of maltose is


present (+)

If the solution has a yellowish coloration, then a moderate amount of maltose


is present (++)

If the solution has an orange or reddish coloration, then a large amount of


maltose is present. (+++)

Prediction:
Make a prediction as to which test tube will have the best starch digestion. Why do you
think this?

Data/Observations:
Test Tube
number

Contents

Lugols Test
Results

Benedicts Test
Results

Conclusion:
Which test tube had the best starch digestion? How do you know? Why do you think
this occurred?

Part 2: Protein digestion by pepsin


Background: Pepsin in the stomach breaks down proteins into smaller polypeptides?
1. Obtain a set of five labeled test tubes
2. Add a small sliver of hard-boiled egg white to each of the five tubes
3. Add the following solutions to the tubes (see Fig 11.10):

TUBE 1: Add 5 ml pepsin solution and 10 drops of water

TUBE 2: Add 5 ml pepsin solution and 10 drops of HCl

TUBE3:Add 5ml pepsin solution and 10 dropsHCl, then place on ice.

TUBE 4: Add 5 ml water and 10 drops HCl

4. Place Tubes 1, 2, and 4 in a 37 C water bath to incubate for 90 min.


5. Examine the egg white following incubation. Note any digestion that occurs to the
egg white (Fig 11.11), and any color changes that may have happened to the
solution.

Prediction:
Which test tube do you think will have the best protein digestion? Why?

Data and Observations


Test Tube

Observations

Part 3: Lipid Digestion


Background: Lipid digestion occurs in the small intestines because of the enzyme
lipase.
In the presence of lipase
Triglycerid + Water Glycerol + Fatty Acids
1. Obtain a set of 3 test tubes
2. Add 3.0 ml heavy cream to each tube
3. Add the following to each tube:

TUBE 1: 5 ml water and a few grains of bile salts

TUBE 2: 5 ml pancreatin solution

TUBE 3: 5 ml pancreatin and a few grains of bile salt

4. Shake tubes vigorously to mix the contents


5. Keep in a water bath at 37 C for 60 minutes. Record the pH every 20 minutes.

Prediction:
Which tube will be the best to digest lipids? Why?

Data and Observations:

Time

Tube 1

Tube 2

Tube 3

Conclusion:
What is necessary for lipid digestion?

Post Lab Questions:


1. What enzyme will further break down maltose? To what will it be broken down?

2. In addition to the mouth, where does starch break down occur?

3. What affect did boiling have on the salivary amylase? Why?

4. What was the control in the amylase experiement? Is it a positive or negative


control?

5. Why would you predict that pepsin would not digest starch?

6. What is the inactive form of pepsin? What cell releases pepsin?

7. Why were test tubes kept specifically at 37 C?

8. What pH change did you observe when fat was digested? Why was this change
observed?

9. What is the role of bile salts in lipid digestion?

10.
List and explain AT LEAST two (2) sources of error in your experimental
procedure.

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